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I Gohin

Publications and source records attributed to I Gohin.

4 recordsLinked to original sources

Analysis of the immune response in sheep efferent lymph during Salmonella abortusovis infection.

The efferent lymph duct of the ovine prescapular lymph node was cannulated, and Salmonella abortusovis (SAO), a specific pathogen for sheep inducing abortion and mortality of newborn lambs, was inoculated by the subcutaneous route in this lymph node drained area. While the prescapular lymph node draining the inoculation site represented an efficient barrier for the vaccinal SAO Rv6 strain spreading, SAO 15/5 virulent bacteria were steadily detected in efferent lymph of infected sheep. The inoculation of the virulent strain of SAO induced a greater increase of the cell output than did the attenuated vaccinal strain, but proportions of blast cells appearing in the efferent lymph were similar in both cases. Flow cytometry analysis showed that B and T cell outputs were both increased during SAO infections, but while T cell subset proportions slightly decreased, B cell percentages significantly rose, and, at the peak response, almost all of the lymphoblast cells were activated B cells. Typical antibody profiles characteristic of a primary immune response were observed, and antibody titres were greater in the efferent lymph of animals inoculated with the virulent strain of SAO. Many of the cytokine mRNAs we investigated were steadily detected by RT-PCR in efferent lymph cells of control sheep, but frequencies of detection of IL-2, IFN gamma, IL-1 beta and TNF alpha mRNAs were augmented in efferent lymph cells following inoculation of both SAO virulent or vaccinal strains. IL-10 and IL-8 mRNAs could only be detected after a SAO inoculation, while detection of IL-4 mRNAs was increased only in efferent lymph cells from SAO virulent strain-infected sheep. The efferent lymph cannulation technique thus appeared a very powerful way to study the in vivo development of the immune response to SAO, in its natural host, the sheep.

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Cellular composition of Corynebacterium pseudotuberculosis pyogranulomas in sheep.

The cellular composition of 46 pyogranulomas experimentally induced with Corynebacterium pseudotuberculosis in sheep was determined by immunohistochemistry. Lesions localized in inoculation sites or draining lymph nodes consisted of macrophage and lymphocyte layers distributed around a necrotic center and surrounded by a fibrous capsule. In immature lesions, T cells of the CD4+ subset predominated, whereas in mature lesions proportions of CD8+ T lymphocytes and cells expressing the gamma/delta chains for the T cell receptor increased. Numerous pyogranuloma cells expressed the interleukin-2 receptor. In addition to these general characteristics, a large individual variability in the proportions of macrophage and T cell subsets was observed for lesions of the same age, in particular for epithelioid macrophages. This heterogeneity suggests a different cellular pattern in relation to the persistence or the elimination of bacteria by the host.

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Humoral immune response to Salmonella abortusovis in sheep: in vitro induction of an antibody synthesis from either sensitized or unprimed lymph node cells.

In vitro culture conditions were determined to induce an anti-Salmonella abortusovis antibody synthesis from lymph node leucocytes of three immunized sheep and two unprimed lambs maintained in culture in the presence of heat-inactivated bacteria for 2 weeks. Humoral immune responses were assessed by enumerating specific antibody-secreting cells using ELISASPOT and by titrating immunoglobulins secreted into culture supernatants using ELISA techniques. Optimal secondary antibody response was observed from cultures performed with fetal calf serum (compared with horse serum) and with an antigen concentration of one to ten bacteria per cell. This kind of antigenic stimulation allowed induction of numerous antibody-secreting cells without adsorption of the secreted antibodies. Maximal numbers of antibody-secreting cells could reach a rate of 1% of the sheep leucocytes initially put into culture. Kinetic profiles of antibody production from boosted lymph node cells were characterized by an ascending phase from the sixth to the twelfth day of culture and then showed a plateau phase until Day 14. Most of the responses were composed of IgM and IgG1 antibodies, traces of IgG2 being detected at the end of experiments. From the twelfth day of antigenic stimulation, the IgM isotype was preferentially expressed with high antigen concentration (100 bacteria per cell), whereas the highest amounts of IgG1 were detected at lower concentration (one to ten bacteria per cell). While anti-Salmonella IgM appeared to be mainly specific for the lipopolysaccharide (LPS) cell wall fraction, some IgG1 recognized other bacterial antigens. Kinetic profiles and magnitudes of primary antibody responses induced in vitro from lamb lymph node cells did not differ from those defined in cultures of sheep boosted leucocytes. But these immune reactions were mainly made up of anti-LPS IgM. Few anti-Salmonella IgG1 were detected from the tenth day of culture. So these in vitro assays allowed induction of antibody synthesis from either in vivo sensitized or unprimed sheep lymph node leucocytes. This methodology would permit achievement of more detailed studies on interactions between Salmonella and lymph node leucocytes, leading to a better understanding of the mechanisms controlling bacterial dissemination through the lymphoid tissue.

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[The lymphatic system and its functioning in sheep].

Cannulation of the afferent and efferent lymph ducts of a lymph node in large domestic species have greatly facilitated the study of the immune system functions in mammals, both in physiological conditions and during immune responses to various pathogens. Summarizing the numerous studies undertaken in the ovine species, this review first presents a description of the structure and composition of the lymph nodes, vessels as well as the lymph fluid they transport. The functioning and the role of each of these immune compartments are also discussed. The characteristics and mechanisms of lymphocyte recirculation between the blood and the lymphatic system are treated in an other chapter. The last part is dedicated to analyzing the modifications observed in the ovine lymphatic system after antigenic stimulation.

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